恒河猴静脉内皮细胞培养的研究

STUDIES ON CULTURE OF MACAQUE'S VENOUS ENDOTHELIAL CELLS IN VITRO

利用胶原酶对恒河猴血管内皮细胞进行消化,以9.92±3.34×10~3个细胞/cm^2的接种率接种于35mm培养皿中原代培养,体外培养7.7±1.82天,细胞增长了7.39±5.04倍,以1:6及1:2比例分别传第一代、第二代共历时13.89±1.36天细胞增长了147.93±88.68倍。对原代及传代细胞进行染色体及第Ⅷ因子相关抗原免疫荧光检查,证实所培养的细胞为内皮细胞。通过检测培养上清中vWF和PGF1α的含量,原代、第一代与第二代之间均无明显差异(P>0.05)。

The macaque's venous endothelial cells were harvested with 0.1% collagenase and then inoculated into the plastic culture plate (diameter 3.5 cm) at a density of 9.92±3.34 × 103 cells/cm2 for primry culture. The primary culture was passaged from diameter 3.5 cm plates to diameter 8.8 cm plaetes and then to another two 8.8 cm plates which was reached on day 13.89±1.36. The splitting rate was 1:6 at first passage and 1:2 at second passage. When culture were terminated, 11.46±2.69 × 106 of avaialbe endothelial celis could be regularly obtai-ned, which was 147.93±88.68 folds as compared with those harvested from host vein. The cultures were confirmed as endothelial cells with the fluorescent linked anti-Ⅷ antigen anti-bodies. The content of both 6-keto-PGF 1α and vWF in the supernatant were detected with ELISA and radioimmunoassay. There were no difference among the primary and subcultures endothelial cells.